Identification of Waitea circinata as a Pathogen of the Moss Bryum argenteum var. argenteum on a Golf Course Fairway.

Waitea circinata Warcup and Talbot (also referred to as W. circinata var. circinata) is an important fungal pathogen of amenity turfgrasses and is especially problematic on Poa annua in putting greens in the late spring or early summer. The pathogen was first identified in 2005 from Japan and has since been seen widely throughout the United States (1,2). On occasion, the pathogen has been observed attacking creeping bentgrass (Agrostis stolonifera) but is typically less virulent. Disease symptoms include prominent yellow rings appearing throughout established turf and moderate leaf necrosis. In the summer of 2012, moss from a section of fairway on a golf course in Edwards, CO was observed to be rapidly killed by a fungal pathogen producing copious amounts of aerial mycelium and appearing similar to Waitea microscopically. Aerial mycelium was transferred to acidified potato dextrose agar (PDA) (1 ml lactic/L). After 1 day at 25°C, mycelia were transferred to PDA. After 2 weeks, plates were covered with white aerial mycelium and separate, spherical, 0.5-mm diameter, salmon-colored sclerotia, which turned dark brown within a few days and were produced submerged throughout the media. Spores were never produced and right-angled branching of mycelia, characteristic of Waitea, was observed in mature cultures. Mycelial plugs were incubated in nutrient broth and DNA was extracted using a MoBio Power Plant DNA extraction kit. Amplification of ribosomal ITS sequences with ITS4 and ITS5 resulted in a 100% identity match with GenBank sequence HM807352, W. circinata var. circinta (3). To demonstrate pathogenicity on Bryum argenteum, unaffected moss from the submitted sample (identified as B. argenteum var. argenteum via 100% sequence identity with the published GenBank sequence GU907062) was removed from the originally submitted sample and placed in separate growth chambers at 95% humidity and 21, 26, and 31°C. An additional experiment employed local B. agenteum plants collected from the URI Kingston, RI campus. Agar plugs from the isolated W. circinata were placed on top of the moss and within 2 days the fungus had caused complete mortality at all three temperatures. The experiment was also undertaken using the same environmental conditions with 5-week-old annual bluegrass (P. annua) and creeping bentgrass cv. A4 grown from seed. Plants were inoculated with infected rye grains at 31, 26, and 21°C. After 1 week, the P. annua plants showed significant mortality at 26 and 31°C with little infection at 21°C and the A. stolonifera plants showed moderate mortality at 26°C and little infection at the other two temperatures. All experiments utilized an additional uninoculated control treatment that showed no moss/turf necrosis or mortality. Experiments were all repeated once and used three replicates per experiment. While moss is not intentionally cultivated on golf courses, it does occur with regularity and often presents itself as a difficult pest to manage. This particular isolate of W. circinata has identical ribosomal and physiological characteristics of the reported P. annua pathogen but can attack one moss species and may be a possible candidate for selective biological control of moss in golf course settings. It is unclear how widespread moss pathogenicity is within W. circinata. References: (1) E. N. Njambere et al. Plant Dis. 95:78 2011. (2) T. Toda et al. Plant Dis. 89:536, 2005. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications, 1990.